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Phosphatidylinositol‐specific phospholipase C From Bacillus cereus : Improved purification, amino acid composition, and amino‐terminal sequence
Author(s) -
Volwerk Johannes J.,
Wetherwax Peter B.,
Evans Loreene M.,
Kuppe Andreas,
Griffith O. Hayes
Publication year - 1989
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.240390311
Subject(s) - bacillus cereus , biochemistry , amino acid , phospholipase c , chromatography , polyacrylamide gel electrophoresis , chemistry , gel electrophoresis , peptide sequence , phospholipase , phosphatidylinositol , ammonium sulfate precipitation , biology , enzyme , size exclusion chromatography , bacteria , genetics , kinase , gene
Phosphatidylinositol‐specific phospholipase C was purified in a 27% yield from the culture medium of Bacillus cereus by a combination of ammonium sulfate precipitation and ion‐exchange and hydrophobic interaction chromatography. The purified enzyme was free of other phospholipase C‐type activities and exhibited a high specific activity of approximately 1,300 units/mg. Amino acid composition analysis and sodium dodecyl sulfate‐polyacrylamide gel electrophoresis indicated a molecular weight of about 35 kDa. The sequence of the first 29 N‐terminal amino acids was also determined.